8PPI

Human inositol 1,4,5-trisphosphate 3-kinase A (IP3K) catalytic domain in complex with beta-D-glucopyranosylmethanol 3,4,1'-trisphosphate/ATP/Mn

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-2
Synchrotron site	ESRF
Beamline	ID23-2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-10-06
Detector	DECTRIS PILATUS3 2M
Wavelength(s)	0.873128
Spacegroup name	C 2 2 21
Unit cell lengths	73.953, 98.124, 193.163
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	96.580 - 1.650
R-factor	0.18636
Rwork	0.185
R-free	0.21405
Structure solution method	FOURIER SYNTHESIS
RMSD bond length	0.005
RMSD bond angle	1.379
Data reduction software	autoPROC
Data scaling software	autoPROC
Phasing software	REFMAC
Refinement software	REFMAC (5.8.0258)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	193.160	1.680
High resolution limit [Å]	1.650	1.650
Rpim	0.024	0.493
Number of reflections	77850	4142
<I/σ(I)>	15.5	1.7
Completeness [%]	91.9	100
Redundancy	12.8	11.6
CC(1/2)	0.999	0.620

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	291	0.81 M sodium citrate, 0.1M Tris pH 8.5 and 0.1 M NaCl. Protein:precipitant ratio 1:1. Protein concentration: 18 mg/ml. Protein buffer: 20 mM Tris pH 7.5, 50 mM ammonium sulfate and 2 mM DTT. Soaking 2h with 1.5 M lithium sulfate, 0.1 M Tris pH 8.5, 5 mM ligand, 3 mM ATP and 3 mM MnCl2.